CN102539395A - Method and device for microscope imaging of a structure of a sample - Google Patents
Method and device for microscope imaging of a structure of a sample Download PDFInfo
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Abstract
The method involves checking presence of a predetermined activation state and detecting changes of corresponding image signals between two test single frames, where presence of the predetermined activation state is found if the detected changes of the image signals collectively exceed a predetermined quantity. A sample structure is imaged onto sensor elements when the predetermined activation state is present. The sensor elements generate respective image signals resulting in a single frame of a sample structure image. An independent claim is also included for a device for light-microscope imaging of a sample structure.
Description
Technical field
The present invention relates to the method and apparatus that is used for the sample structure micro-imaging according to claim 1 or 10 preambles.
Background technology
Exploitation in recent years has some optical microphotograph formation methods; With these methods can based on some single markings, particularly fluorescence molecule continuously, at random locating and displaying goes out sample structure, these sample structures are less than the resolution limit under the microscopical diffraction conditions of traditional optical.Such method for example is described in WO 2006/127692 A2; DE 10 2,006 021 317 B3; WO 2007/128434 A1, US 2009/0134342 A1; DE 10 2,008 024 568 A1; " Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM) " Nature Methods 3,793-796 (2006), M.J.Rust, M.Bates, X.Zhuang; " Resolution of Lambda/10 in fluorescence microscopy using fast single molecule photo-switching ", Geisler C.et al, Appl.Phys.A, 88, among the 223-226 (2007).This new microtechnic branch also is called as the location microtechnic.Applied method for example in document with symbol (F) PALM ((fluorescence) photoactivate location micro-); PALMIRA (having the independent PALM that obtains that carries out), GSD (IM) (ground state loss unimolecule returns) is micro-) perhaps (F) STORM ((fluorescence) optics reconstruct is micro-at random) is open.
New method is normally prepared sample structure to be formed images in advance with some marks, and these marks have two diacritic states, just " bright " state and " secretly " state.If for example use fluorescent colorant to serve as a mark, then bright state is that fluorescigenic state of ability and dark state are can not fluorescigenic state.For resolution imaging sample structure, the sub-fraction of mark repeatedly is converted into bright state with the resolution limit that is higher than traditional imaging optical system.This part is called as active part below.At this, the density that forms mark that should " activity " part is selected like this, but make bright state and thus the average headway of the adjacent marker in the state of optical microphotograph imaging greater than the resolution limit of imaging optical system.The mark that forms active part is imaged on the spatial discrimination device of sensor element, makes that the size of hot spot is confirmed by the resolution limit of lens combination by the light distribution of each marker detection to one hot spot form.
If it is in this wise little to be included in the density of the mark in the active part, make the average headway of foregoing mark surpass by the predetermined minimum spacing of the resolution limit of optical microphotograph imaging device, then be to be in desirable active state.This active state guaranteed, accurately comes from the mark of an activation by each of the detected hot spot of arrangement of sensor elements.
Described before method is by explanation visually in Fig. 1.The sequence of a raw data-single image has been shown in the left part of Fig. 1, and said single image is by with 1 ..., N numbers one by one.Be illustrated in hot spot in these raw data-single images respectively from the mark part of an activation; Said part is in the desirable active state; That is to say; Its mark density is so for a short time, makes the average headway guaranteed these marks greater than by the predetermined minimum spacing of the resolution limit of optical microphotograph imaging device.
An overall diagram has been shown in the upper right portion of Fig. 1, and this overall diagram is by raw data-single image 1 ..., N is formed by stacking.This overall diagram has a light fuzzy, that spatially do not differentiated and distributes, and this light is distributed in and does not describe out sample structure to be formed images on the details.
An overall diagram has been shown in the bottom left section of Fig. 1, and this overall diagram is formed by stacking the photodistributed barycenter of being determined raw data-single image.This overall diagram has obviously higher resolution, make can be well on details-in this example with its three concentric these sample structures of annulus-identification.
In order to quantize accessible bearing accuracy in said method, consult Fig. 2, the fact of case that is illustrated among Fig. 1 is schematically explained in Fig. 2 by pure once more.The sequence of a raw data-single image has been shown in Fig. 2 top.Sample structure to be formed images, that be made up of some donuts is drawn with dotted line in Fig. 2 and with reference number 2 marks.
Each mark of active part produces light and distributes 4, and these photodistributed sizes provide and in Fig. 2, use Δ x by the resolution capabilities of imaging optical device
AbbIdentify.As in Fig. 2, schematically showing ground, said mark has less than said size, delta x each other
AbbAverage headway, make these light distribute and 4 do not superpose.
Light has been shown in Fig. 2 bottom has distributed 6, these light distribute and confirm to be drawn by light distribution 4 through barycenter.Attainable bearing accuracy is by the size, delta x of light distribution 6
SPProvide.For bearing accuracy Δ x
SPBe applicable to:
Wherein, n representes according to the light 4 detected photon numbers that distribute.Positional precision Δ x
SPTypically be arranged in about scope of 10 to 50nm.
Before taking raw data-single image, set up aforesaid active state.Which kind of initial state to regulate said active state based on, be different between method and the method.Can be divided into two methods in principle, that is, and a method and a method that is called " from top to bottom " that is called " from bottom to top ".Underlined at first being in the dark state in the method from bottom to top.For the generation of the required activity mark in high-resolution location part by corresponding activation, for example the radiation by activation light causes.This for example is the situation in (F) PALM method.On the contrary, but in top-down method underlinedly be at first that it is bright, in state of optical microphotograph imaging just.In order to produce activity mark's part, the major part of these marks is converted in its dark state subsequently, for example the radiation through light once more.At this, these marks can be so-called " convertible " marks, for example convertible protein.These marks also can be some conventional fluorescent colorants of confirming, and these colorants change long-term dark state, for example triplet (and being inconvertible mark thus in itself) into through the radiation of excitation line.For example in GSD (IM) method, work in this way.
All compare in method from bottom to top and the top-down method difficulty be, regulate predetermined active state with reproducible mode, this active state can realize the high-resolution location of each mark.So far especially can not confirm whether this predetermined active state exists with simple mode.Therefore can occur usually, too many mark is in its bright state, makes the light that in raw data-single image, is produced by mark distribute spatially inseparable each other.Make like this to the optical microphotograph imaging of sample structure, especially to determining that when can begin substantial measurement causes difficulty; Just when in bright state, only also in fact there is a small amount of mark, makes the light distribution that produces by mark have the space distribution that can realize photodistributed separable detection with sufficiently high probability.
Summary of the invention
Task of the present invention is; To improving according to method claim 1 preamble, that be used for optical microphotograph imaging sample structure; Make simply to reach and confirm that accurately whether have predetermined active state, mark has enough big average headway each other in this state.
The present invention has solved this task thus according to the characteristic of claim 1; Promptly; Check the existence of predetermined active state, its mode is to produce at least two single width checking images at a distance of a time interval ground; At least distinguish the variation of detected image signal between two single width checking images for the part of sensor element, and when the variation of detected picture signal has surpassed predetermined size on the whole, confirm to exist predetermined active state.
The present invention based on understanding be; Can spatially detect in the state of the light signal that is sent by mark at one, the time of said signal changes at it and changes greater than the time of light signal in a situation about not detected separated from one anotherly generally separated from one anotherly.In first kind of situation, as if can be identified in the flash of light in the micro-image, this flash of light typically is used for the existence of the desirable active state of mark thus.This has realized thus; But Random assignment always ground with some marks from its state (for example passing through photobleaching) bright, optical microphotograph imaging just transform to its dark state and other mark from its dark state transformation to its bright state, these are marked at it and are converted to the photon of also launching some before the dark state once more in bright state.
The present invention utilizes foregoing phenomenon, is used for check and whether has predetermined active state.This so realizes, promptly takes at least two single width checking images at a distance of a time interval ground and the variation of the picture signal that produced by sensor element is confirmed as and is used for foregoing flash of light how strong standard is arranged.If the space density that is marked in the bright state is big like this, make the light signal that sends by each mark superpose, then flasher characteristic is strong slightly.More multiple labeling sends on its light signal to one and the same sensor element, then flasher characteristic in micro-image more a little less than.The time average of the light signal that has just carried out in the case each sensor element is received.
The present invention has stipulated, is used to the active standard of glistening by the change-detection between two single width checking images taking at a distance of a time interval ground, and when can begin the substance imaging to sample structure according to its decision.
Preferably, according to the change of detected picture signal confirm two single width checking images image signals corresponding each other temporal correlation and when this temporal correlation is lower than predetermined relevance threshold, confirm to exist predetermined active state.The temporal correlation of two front and back single images in succession is one and is suitable for confirming the active measurement parameter that has desirable active state thus that reaches of flash of light.Therefore, the temporal correlation between two single width checking images is more little, and the activity of then glistening is high more.
In a favourable further improvement according to the method for the invention, stipulate; When detected picture signal is arranged in one first codomain; For the picture signal of detected two single width checking images disposes one first value respectively; And when detected picture signal is arranged in second codomain with first codomain complementation, for the picture signal of detected two single width checking images disposes second value different with first value respectively.For example can be with picture signal Configuration Values 0 less than a predetermined threshold value, and will be equal to or greater than the picture signal Configuration Values 1 of this threshold value.Therefore, draw the data recording that some only comprise value 1 and 0 respectively, and be suitable for for example active thus particularly well through confirming that temporal correlation detects flash of light by these two single width checking images.
The time interval that produces two single width checking images apart is preferably by such adjusting, but makes this time interval approximate to be marked at 1 times to 3 times of mean residence time in the state of optical microphotograph imaging.As long as the time interval moves in this scope, then having guaranteed when having predetermined active state, to have sufficiently high reaching thus can be active according to two detected flashes of light of single width checking image.
Preferably; Exist under the precondition of predetermined active state; Active state is check when reducing the time interval that produces two single width checking images apart with being repeated; Variation up to detected picture signal no longer surpasses a predetermined size on the whole at it, and under the situation of considering the time interval, regulates the time shutter of sensor element, and the variation of detected picture signal no longer surpasses predetermined size on the whole at it in this time interval.The preferred configuration of said method has been stipulated, can shorten the time interval that produces two single width checking images apart step by step based on desirable active state, up to no longer there being active state.Can draw a particularly advantageous time shutter through this embodiment, i.e. short as far as possible time, but should the time also long enough so that can detect the active state of existence reliably.
Usually, like the illustrated ground of beginning part, take an image sequence that constitutes by a plurality of single images, so that can form images sample structure.In the case, before taking each single image, carry out that is to say, before taking each single image, check the existence of predetermined active state according to checking procedure of the present invention.Also have advantage at this, the single width checking image that will during checking procedure, take (form to revise in case of necessity) is considered as the single image of image sequence, exists as long as in this checking procedure, confirmed predetermined active state.Also can be only with one in these single width checking images single image as image sequence.
In addition, the present invention has stipulated device characteristic, that be used for optical microphotograph imaging sample structure with explanation in claim 10.
The present invention is specified according to accompanying drawing below.Wherein show:
Description of drawings
The sequence of a raw data-single image of Fig. 1 and the overall diagram that is combined into thus before definite at barycenter and afterwards;
Synoptic diagram after image sequence among Fig. 2 Fig. 1 reached before barycenter is confirmed;
Fig. 3 is used to implement the structure of optical microscope according to the method for the invention;
Fig. 4 is used for according to an embodiment process flow diagram according to the inventive method being described; And
Fig. 5 reaches two front and back raw data-single images in succession in the situation that has predetermined active state respectively in the situation that does not have predetermined active state.
Embodiment
Fig. 3 shows an optical microscope 20 as embodiment, and this optical microscope is suitable for implementing the method that is used for a sample structure of optical microphotograph imaging according to of the present invention.
The light that is sent by this sample structure 34 passes object lens 30 and incides on the dichroic mirror 32.This dichroic mirror 32 is by structure like this, makes light that its reflection is sent by sample structure 34 and like this towards lens 38 orientations, but on the photodetector 40 of these lens with said smooth pack to a spatial discrimination.This photodetector 40 is made up of the device of a matrix form of sensor element, for example CCD element.Each of these sensor elements will convert electrical picture signal to by the light of its reception.These picture signals are outputed on the control module 42 subsequently.Control module 42 is controlled each microscope assemblies and is especially had the function that the picture signal that is received by photodetector 40 is handled.
When use is illustrated in the optical microscope 20 among Fig. 3, combine Fig. 1 sample structure 34 that forms images like beginning saidly.Just, take the sequence of a raw data-single image, for these images other mark active part that forms images respectively.Subsequently, in image analysis process, control module 42 is confirmed photodistributed barycenter in each raw data-single image, and these barycenter represent to be in the mark in the bright state.The photodistributed barycenter of being tried to achieve by raw data-single image is combined into a high-resolution overall diagram subsequently.
Stipulate according to the method for the invention; Whether check exists for the high-resolution optical microphotograph necessary active state that forms images before producing each raw data-single image; That is to say; Whether the mark that is included in each activity mark part has an average headway each other, and this average headway is greater than by the predetermined minimum spacing of the resolution limit of optical microphotograph imaging device.This combines the process flow diagram of Fig. 4 to explain below.
The process that is illustrated among Fig. 4 begins with step S1.In step S2, operation variable p is by assignment 1, and raw data-single image is numbered by this operation variable p seriatim.
Follow circulation step S2, that constitute by step S3 to S6 be used to produce a desirable N single image (p=1 ..., sequence N).Therefore, when operation for the first time is used for the step S3 (p=1) of conduct first raw data-single image to be produced, but the part of mark is converted in the state of optical microphotograph imaging, so that produce one first activity mark's part.
The present invention stipulated in step S4, detects the mark that is included in this first activity mark part and whether has an average headway each other, and this average headway is greater than by the predetermined minimum spacing of the resolution limit of optical microphotograph imaging device.Also can be specified at last according to detection of the present invention.Exist if the detection in step S4 draws predetermined active state, then this process is proceeded with step S5 according to Fig. 4, in step S5, produces p raw data-single image.Then in step S6, move variable p added value 1.On the contrary; If in step S4, draw; Also do not have predetermined active state, then this process is jumped and is returned step S3, in this step S3, attempts again; But make mark transitions in the state of its optical microphotograph imaging, so that set up desirable active state with desirable space density.The circulation that is made up of step S3 and S4 is moved so for a long time, and the detection in step S4 draws, and has predetermined active state.
In step S7, all check at every turn, whether move the total N that variable p has reached single image.If also do not reach, then this process is jumped and is returned step S3.In case operation variable p has reached value N, that is to say the desirable number N that has produced single image, the process that then is illustrated among Fig. 4 finishes.
As start saidly, must in raw data-single image, confirm barycenter, so that obtain having desirable high-resolution overall diagram.In the present embodiment, barycenter just is determined after can in step S4, being determined or being illustrated in the process end among Fig. 4.
Be described in detail in the detection of the active state of carrying out among the step S3 below.
In order in step S3, to detect active state, at first each other at a distance of time interval ground, take two single width checking images with raw data-single image form.In the present embodiment, this time interval provided through the time shutter, and photodetector 40 needs the time shutter to be used to produce picture signal.This means that photodetector 40 directly begins to produce second raw data-single image after producing first raw data-single image.It is active to detect flash of light by these two single width checking images subsequently, so that confirm whether desirable active state exists.
Flash of light is active to be detected through the temporal correlation of confirming two single width checking images in the present embodiment.This for example carries out according to following relational expression:
K representes relevance parameter in this relational expression, r
IjBe used for checking image that two single width checking images at first take and represent the signal value of the picture signal of such sensor element, said sensor element is positioned at i capable and j row, wherein i=1 in the layout that forms sensor element photodetector 40, matrix form; 2; ..., n and j=1,2; ..., m and n are the sum and the sum of m for being listed as of row.Correspondingly,
checking image of taking after a while of being used for two single width checking images representes to be positioned at the signal value of picture signal of the sensor element of the capable and j row of i.
The relational expression of explanation shows before, the signal value of two single width checking images is multiplied each other each other and subsequently with the product addition of all the sensors element for each sensor element.That obtain like this and by normalization, its mode is, it is divided by one the quadratic sum of signal value in two single width checking images, and the relevance parameter k that tries to achieve like this gets a value between 0 and 1, just 0≤k≤1 subsequently.If this value k value of being similar to 0, then the temporal correlation between two single width checking images is little, that is to say between two single width checking images significant change to occur.This means that also there is desirable active state thus in active high the reaching of flash of light.
On the contrary, if this relevance parameter k value of being similar to 1, then temporal correlation is big, and between two single width checking images, little variation only occurs.This means that also flash of light is active low.In the case, but it is still too high and must further reduce to be in the space density of the mark in the state of optical microphotograph imaging, so that reach desirable bearing accuracy.
In order to draw the decision whether predetermined active state exists, confirm the threshold value between 0 and 1 in advance.If relevance parameter k then draws less than this threshold value thus, the flash of light activity is sufficiently high and has predetermined active state thus.On the contrary, if not this situation, but then should further reduce the space density of the mark in the optical microphotograph image formation state.
In the present embodiment, under the situation of considering all sensor elements, detect relevance parameter k.But, also can only consider the part of sensor element in order to confirm relevance parameter k.
The described before relational expression that is used for confirming relevance parameter k is based on this, that is, a sensor element that does not receive light is exported the picture signal with value 0.But if possible there is ground unrest, that is to say that each sensor element exports a picture signal that has greater than 0 value in the case, then can in above-mentioned formula, consider this signal, so that remove by average ground unrest.
Also can whether come signal value r according to it respectively greater than a predetermined threshold value
IjWith
Configuration Values 1 perhaps is worth 0.From two single width checking images, only had the data recording of value 0 and 1 respectively through such measure, simplified confirming of relevance parameter k like this.
The threshold value that can compare with the relevance parameter k that tries to achieve is preferably confirmed according to permissible error to be defined in individual cases.At this, referring to for " error " is not from an independent mark but from the photodistributed detection of a plurality of marks, but these marks to each other apart from less than the predetermined minimum spacing of resolution limit by the imaging device of optical microphotograph.
Some real example have been shown in Fig. 5, and these examples have been explained the influence of mark density to raw data-single image with figure.In these examples, the microtubule of PtK2 cell is formed images by optical microphotograph ground under the situation painted with Rh6G.Show two front and back raw data-single images in succession respectively.
In the raw data-single image on two tops of Fig. 5, but the mark density in the state of optical microphotograph imaging is too high, that is to say not have desirable active state.Therefore, these two raw data-single images of taking are successively only distinguished seldom each other.
On the contrary, in the raw data-single image of two bottoms of Fig. 5, but the mark density in the state of optical microphotograph imaging is in this wise little, makes to have desirable active state.Like the shown ground of raw data-single image of taking successively referring to these two, in this active state, can detect each mark.Obviously visible in addition marks, these are marked among of two raw data-single images launches light signal, and is can not detected (vice versa) mostly in other raw data-single image.This means that flash of light is active high when taking these raw data-single image.
Claims (10)
1. method that is used for optical microphotograph imaging sample structure (34), wherein:
Prepare said sample structure (34) with mark, but said mark can be converted to the state of optical microphotograph imaging,
Produce activity mark's part, but the part of wherein said mark be converted in the state of this optical microphotograph imaging,
When having predetermined active state, this sample structure (34) is imaged on the device (40) of sensor element, in said active state; The mark that is included in activity mark's part has average headway each other; This average headway is greater than by the predetermined minimum spacing of the resolution limit of optical microphotograph imaging device, and these sensor elements produce picture signal respectively, wherein; These picture signals provide the single image of sample structure (34) with its integral body
It is characterized in that; Check the existence of this predetermined active state; Wherein produce at least two single width checking images at a distance of a time interval ground; At least for the variation of detected image signal between two single width checking images respectively of the part of sensor element, and in the variation of detected picture signal when it surpasses predetermined size on the whole, confirming should predetermined active state existence.
2. method according to claim 1; It is characterized in that; Confirm according to the variation of detected picture signal two single width checking images picture signal corresponding each other temporal correlation and when this temporal correlation was lower than predetermined relevance threshold, confirming should predetermined active state existence.
3. method according to claim 2 is characterized in that, this temporal correlation is confirmed according to following relational expression:
Wherein,
K representes a relevance parameter,
r
IjBe used for checking image that two single width checking images at first take and represent the signal value of the picture signal of such sensor element, said sensor element is positioned at i capable and j row, wherein i=1 in an arrangement of sensor elements (40) with the capable matrix form with the m row of n; 2 ..., n and j=1; 2; ..., m, and
The checking image of taking after a while that
is used for two single width checking images is represented a signal value that is arranged on the picture signal of the sensor element in the capable and j row of i.
4. according to the described method of one of aforementioned claim; It is characterized in that; When detected picture signal is arranged in first codomain; The detected picture signal of two single width checking images is configured one first value respectively, and when detected picture signal was arranged in complementary second codomain of this first codomain, the detected picture signal of two single width checking images was configured second value different with first value respectively.
5. according to the described method of one of aforementioned claim, it is characterized in that, regulate to produce the time interval of two single width checking images, but make this large interval approximate to be marked at 1 times to 3 times of mean residence time in the state of this optical microphotograph imaging.
6. method according to claim 5 is characterized in that, regulates the time shutter of sensor element, but makes it approximate the mean residence time in the state that is marked at this optical microphotograph imaging greatly.
7. according to the described method of one of aforementioned claim; It is characterized in that; Under the precondition that predetermined active state exists; This active state of duplicate test under the situation about reducing in the time interval that produces two single width checking images, up to the variation of detected picture signal till it no longer surpasses predetermined size on the whole, and; Under the situation of considering the time interval, regulate the time shutter of sensor element, the variation of detected picture signal no longer surpasses this predetermined size on the whole at it in this time interval.
8. according to the described method of one of aforementioned claim, it is characterized in that the image sequence of taking single image reaches the existence of before taking each single image, checking predetermined active state.
9. method according to claim 8 is characterized in that, is determined when existing in this predetermined active state, and in two single width checking images at least one is considered as the single image of image sequence.
10. a device that is used for optical microphotograph imaging sample structure has the mechanism that is suitable for preparing with mark said sample structure, but said mark can be converted in the state of optical microphotograph imaging,
Be suitable for producing the mechanism of activity mark's part, but wherein the part of mark is converted in the state of this optical microphotograph imaging,
Be suitable for when having predetermined active state sample structure is imaged on the mechanism on the layout of a sensor element; In said active state, the mark that is included in this activity mark part has average headway each other, and this average headway is greater than by the predetermined minimum spacing of the resolution limit of optical microphotograph imaging device; Said sensor element produces picture signal respectively; Wherein, these picture signals draw the single image of sample structure with its integral body
It is characterized in that being suitable for checking the mechanism of the existence of this predetermined active state; Wherein produce at least two single width checking images at a distance of a time interval ground; At least distinguish the variation of detected image signal between two single width checking images for the part of sensor element; And when it surpassed predetermined size on the whole, confirming should predetermined active state existence in the variation of detected picture signal.
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| DE102010038265.5 | 2010-10-19 | ||
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| DE102011001091.2 | 2011-03-04 | ||
| DE102011001091.2A DE102011001091C5 (en) | 2010-10-19 | 2011-03-04 | Method and device for microscopic image recording of a sample structure |
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| CN102539395A true CN102539395A (en) | 2012-07-04 |
| CN102539395B CN102539395B (en) | 2016-05-04 |
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| CN201110317596.4A Expired - Fee Related CN102539395B (en) | 2010-10-19 | 2011-10-18 | For the method and apparatus of sample structure micro-imaging |
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| US (1) | US8922638B2 (en) |
| EP (1) | EP2444792B1 (en) |
| JP (1) | JP5891000B2 (en) |
| CN (1) | CN102539395B (en) |
| DE (1) | DE102011001091C5 (en) |
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| DE102012205032A1 (en) * | 2012-03-28 | 2013-10-02 | Carl Zeiss Microscopy Gmbh | High resolution fluorescence microscopy |
| JP7058837B2 (en) * | 2015-11-17 | 2022-04-25 | 韓國科學技術院 | Sample characteristic detector using chaotic wave sensor |
| GB201704275D0 (en) * | 2017-03-17 | 2017-05-03 | Science And Tech Facilities Council | Super-resolution microscopy |
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| US20080182336A1 (en) * | 2006-08-07 | 2008-07-31 | President And Fellows Of Harvard College | Sub-diffraction limit image resolution and other imaging techniques |
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| JP4515899B2 (en) * | 2004-12-17 | 2010-08-04 | オリンパス株式会社 | Microscope image processing method, microscope image processing program, and microscope system |
| EP2453239B1 (en) * | 2005-05-23 | 2017-04-26 | Harald F. Hess | Optical microscopy with transformable optical labels |
| US7776613B2 (en) * | 2006-08-07 | 2010-08-17 | President And Fellows Of Harvard College | Sub-diffraction image resolution and other imaging techniques |
| DE102008021641A1 (en) * | 2008-04-30 | 2009-11-05 | Carl Zeiss Microlmaging Gmbh | Resolution Enhanced Luminescence Microscopy |
| JP5787257B2 (en) * | 2008-05-21 | 2015-09-30 | マックス−プランク−ゲゼルシャフト ツル フォルデルング デル ヴィッゼンシャフテン イー．ヴイ． | High spatial resolution imaging of target structures in a sample |
| DE102008024568A1 (en) | 2008-05-21 | 2009-12-03 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Spatial resolution imaging of structure of interest in specimen involves marking specimen structure, imaging the specimen, exposing the specimen to light, registering the fluorescent light and determining position of molecules of substance |
| EP2438555B1 (en) | 2009-06-02 | 2017-03-08 | Sofast GmbH | Superresolution optical fluctuation imaging (sofi) |
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2011
- 2011-03-04 DE DE102011001091.2A patent/DE102011001091C5/en not_active Expired - Fee Related
- 2011-10-18 CN CN201110317596.4A patent/CN102539395B/en not_active Expired - Fee Related
- 2011-10-18 JP JP2011228719A patent/JP5891000B2/en active Active
- 2011-10-18 US US13/275,393 patent/US8922638B2/en not_active Expired - Fee Related
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Also Published As
| Publication number | Publication date |
|---|---|
| US20120092479A1 (en) | 2012-04-19 |
| JP5891000B2 (en) | 2016-03-22 |
| DE102011001091A1 (en) | 2012-04-19 |
| US8922638B2 (en) | 2014-12-30 |
| JP2012103691A (en) | 2012-05-31 |
| DE102011001091C5 (en) | 2022-07-14 |
| CN102539395B (en) | 2016-05-04 |
| DE102011001091B4 (en) | 2017-09-14 |
| EP2444792B1 (en) | 2018-10-10 |
| EP2444792A1 (en) | 2012-04-25 |
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